Differential analysis of the cytochrome p450 acaricide-resistance genes in Panonychus citri (Trombidiformes: Tetranychidae) strains

Authors

  • Gaofei Jiang Key Laboratory of Entomology and Pest Control Engineering, College of Plant Protection, Southwest University, Chongqing 400716, P.R. China Citrus Research Institute, Chinese Academy of Agricultural Sciences, National Engineering Research Center for Citrus, Chongqing 400712, P.R. China
  • Yunfei Zhang College of Horticulture and Landscape Architecture, Key Laboratory of Horticulture Science for Southern Mountainous Regions, Ministry of Education, Southwest University, Chongqing 400715, P.R. China Citrus Research Institute, Chinese Academy of Agricultural Sciences, National Engineering Research Center for Citrus, Chongqing 400712, P.R. China
  • Fei Chen Key Laboratory of Entomology and Pest Control Engineering, College of Plant Protection, Southwest University, Chongqing 400716, P.R. China Citrus Research Institute, Chinese Academy of Agricultural Sciences, National Engineering Research Center for Citrus, Chongqing 400712, P.R. China
  • Junli Li College of Horticulture and Landscape Architecture, Key Laboratory of Horticulture Science for Southern Mountainous Regions, Ministry of Education, Southwest University, Chongqing 400715, P.R. China Citrus Research Institute, Chinese Academy of Agricultural Sciences, National Engineering Research Center for Citrus, Chongqing 400712, P.R. China
  • Xiaojiao Li College of Horticulture and Landscape Architecture, Key Laboratory of Horticulture Science for Southern Mountainous Regions, Ministry of Education, Southwest University, Chongqing 400715, P.R. China Citrus Research Institute, Chinese Academy of Agricultural Sciences, National Engineering Research Center for Citrus, Chongqing 400712, P.R. China
  • Jiansu Yue College of Horticulture and Landscape Architecture, Key Laboratory of Horticulture Science for Southern Mountainous Regions, Ministry of Education, Southwest University, Chongqing 400715, P.R. China Citrus Research Institute, Chinese Academy of Agricultural Sciences, National Engineering Research Center for Citrus, Chongqing 400712, P.R. China
  • Haoqiang Liu Citrus Research Institute, Chinese Academy of Agricultural Sciences, National Engineering Research Center for Citrus, Chongqing 400712, P.R. China
  • Hongjun Li Citrus Research Institute, Chinese Academy of Agricultural Sciences, National Engineering Research Center for Citrus, Chongqing 400712, P.R. China
  • Chun Ran Key Laboratory of Entomology and Pest Control Engineering, College of Plant Protection, Southwest University, Chongqing 400716, P.R. China College of Horticulture and Landscape Architecture, Key Laboratory of Horticulture Science for Southern Mountainous Regions, Ministry of Education, Southwest University, Chongqing 400715, P.R. China Citrus Research Institute, Chinese Academy of Agricultural Sciences, National Engineering Research Center for Citrus, Chongqing 400712, P.R. China

Keywords:

citrus red mite, differential analysis, cytochrome P450, hexythiazox resistance, miticide resistance, RT-PCR analysis

Abstract

The citrus red mite, Panonychus citri (McGregor) (Trombidiformes: Tetranychidae), a highly destructive pest in citrus groves around the world, has developed resistance to many registered acaricides. Hexythiazox is a selective miticide that has been widely used to control citrus mites in a variety of crops. Forty-six cytochrome P450 mono-oxygenase genes related to general pesticide resistance in other insect species were obtained from the transcriptomes of the hexythiazox-resistant (RR) and hexythiazox-susceptible (SS) strains of P. citri and divided into 4 clans, 15 families and 24 subfamilies. Sequence analyses of each CYP resulted in detection of 3 mutationsin the CYP307A1 gene (841-A to C, 1395-T to C, 1491-T to C) that differed between the 2 strains. Only the change at an amino acid position (278-lysine to glutamine) resulted in a sense mutation. One SNP site was also detected in CYP381A2 (40-A to T) causing a sense amino acid mutation (14-threonine to serine). Seven of these P450s belonged to the CYP2 clan, CYP3 clan and CYP4 clan based on digital gene expression (DGE) library sequencing with a |log2 ratio| value greater than 2, but there were no significant differences revealed by qRT-PCR analysis. This study provides essential information for future research on the hexythiazox-resistance mechanism of P. citri. More methods are needed to further elucidate the molecular mechanisms of resistance to hexythiazox in P. citri.

 

Resumen

El ácaro rojo de los cítricos, Panonychus citri (McGregor), es una plaga muy destructiva en plantaciones de cítricos por todo el mundo y ha desarrollado resistencia a muchos acaricidas registrados. Hexitiazox es un acaricida selectivo que ha sido ampliamente utilizado para controlar los ácaros de cítricos en una variedad de cultivos. Se obtuvieron cuarenta y seis genes del citocromo P450 monooxigenasa relacionados con la resistencia general de pesticidas en especies a partir de los transcriptomes de una cepa de P. citri resistente al hexitiazox (RR) y una cepa de P. citri susceptible al hexitiazox (SS) y fueron divididos en 4 clanes, 15 familias y 24 subfamilias. Los análisis de secuencias de cada CYP resultó en la detección de 3 mutaciones en el gen CYP307A1 (841 - A a C, 1395 - T a C, 1491 - T a C) que difieren entre las 2 cepas. Sólo el primer cambio resultó en una mutación de sentido en una posición de aminoácido (278-lisina a glutamina). También, un sitio de SNP fue detectado en CYP381A2 (40-A a T) que causa una mutación de sentido de aminoácidos (14-treonina a serina). Siete de estos P450 pertenecían al clan CYP2, CYP3 y CYP4 basado en la secuenciación de biblioteca de la expresión de genes digital (EGD) con un valor | log2 ratio| superior de 2, pero no hubo diferencias significativas reveladas por el análisis de RT -PCR. Este estudio provee información esencial para investigaciones en la futura sobre el mecanismo de resistencia para hexitiazox por P. citri. Más métodos para dilucidar los mecanismos moleculares de resistencia a hexitiazox en P. citri son necesarios.

 

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Issue

Vol. 98, No. 1 (March 2015)

Section

Research Papers

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